Product: Phospho-PDPK1 (Ser241) Antibody
Catalog: AF3018
Description: Rabbit polyclonal antibody to Phospho-PDPK1 (Ser241)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Dog, Chicken
Mol.Wt.: 63kDa; 63kD(Calculated).
Uniprot: O15530
RRID: AB_2834425

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
Phospho-PDPK1 (Ser241) Antibody detects endogenous levels of PDPK1 only when phosphorylated at Serine 241.
RRID:
AB_2834425
Cite Format: Affinity Biosciences Cat# AF3018, RRID:AB_2834425.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

3 phosphoinositide dependent protein kinase 1; 3-phosphoinositide-dependent protein kinase 1; hPDK 1; hPDK1; MGC20087; MGC35290; OTTHUMP00000159109; OTTHUMP00000159110; OTTHUMP00000174525; PDK1; Pdpk1; PDPK1_HUMAN; PDPK2; PDPK2P; PkB kinase; PkB kinase like gene 1; PkB like 1; PRO0461; Protein kinase;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O15530 PDPK1_HUMAN:

Appears to be expressed ubiquitously. The Tyr-9 phosphorylated form is markedly increased in diseased tissue compared with normal tissue from lung, liver, colon and breast.

Description:
PDK1 an AGC kinase of the PKB family that contains a PH domain. Involved in a wide variety of processes including cell proliferation, differentiation and apoptosis. Autophosphorylation in the activation loop is necessary for activity.
Sequence:
MARTTSQLYDAVPIQSSVVLCSCPSPSMVRTQTESSTPPGIPGGSRQGPAMDGTAAEPRPGAGSLQHAQPPPQPRKKRPEDFKFGKILGEGSFSTVVLARELATSREYAIKILEKRHIIKENKVPYVTRERDVMSRLDHPFFVKLYFTFQDDEKLYFGLSYAKNGELLKYIRKIGSFDETCTRFYTAEIVSALEYLHGKGIIHRDLKPENILLNEDMHIQITDFGTAKVLSPESKQARANSFVGTAQYVSPELLTEKSACKSSDLWALGCIIYQLVAGLPPFRAGNEYLIFQKIIKLEYDFPEKFFPKARDLVEKLLVLDATKRLGCEEMEGYGPLKAHPFFESVTWENLHQQTPPKLTAYLPAMSEDDEDCYGNYDNLLSQFGCMQVSSSSSSHSLSASDTGLPQRSGSNIEQYIHDLDSNSFELDLQFSEDEKRLLLEKQAGGNPWHQFVENNLILKMGPVDKRKGLFARRRQLLLTEGPHLYYVDPVNKVLKGEIPWSQELRPEAKNFKTFFVHTPNRTYYLMDPSGNAHKWCRKIQEVWRQRYQSHPDAAVQ

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Chicken
100
Xenopus
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O15530 As Substrate

Site PTM Type Enzyme
Y9 Phosphorylation P07949 (RET) , P12931 (SRC)
S22 Phosphorylation
S25 Phosphorylation O15530 (PDPK1)
T33 Phosphorylation P12931 (SRC)
S36 Phosphorylation
T37 Phosphorylation
S64 Phosphorylation
K83 Ubiquitination
S92 Phosphorylation
K111 Ubiquitination
K123 Ubiquitination
T148 Phosphorylation
Y156 Phosphorylation
S160 Phosphorylation
Y161 Phosphorylation
S176 Phosphorylation
T180 Phosphorylation
S231 Phosphorylation
S234 Phosphorylation
K235 Ubiquitination
S241 Phosphorylation O15530 (PDPK1)
T245 Phosphorylation
Y248 Phosphorylation
T255 Phosphorylation
K257 Ubiquitination
Y288 Phosphorylation
K304 Acetylation
K304 Ubiquitination
K315 Ubiquitination
K323 Ubiquitination
K337 Ubiquitination
T354 Phosphorylation Q14680 (MELK)
S366 Phosphorylation
Y373 Phosphorylation P08069 (IGF1R) , P12931 (SRC)
Y376 Phosphorylation P08069 (IGF1R) , P12931 (SRC)
S393 Phosphorylation O15530 (PDPK1)
S394 Phosphorylation Q99683 (MAP3K5)
S396 Phosphorylation O15530 (PDPK1)
S398 Phosphorylation Q99683 (MAP3K5)
S410 Phosphorylation O15530 (PDPK1)
K441 Ubiquitination
Y485 Phosphorylation P12931 (SRC)
Y486 Phosphorylation
K495 Ubiquitination
S501 Phosphorylation Q04759 (PRKCQ)
K509 Ubiquitination
T513 Phosphorylation O15530 (PDPK1) , P12931 (SRC)
S529 Phosphorylation Q04759 (PRKCQ) , O15530 (PDPK1)
K534 Acetylation

PTMs - O15530 As Enzyme

Substrate Site Source
O00141 (SGK1) T256 Uniprot
O00141 (SGK1) S422 Uniprot
O14920 (IKBKB) S181 Uniprot
O15530 (PDPK1) S25 Uniprot
O15530-1 (PDPK1) T33 Uniprot
O15530-4 (PDPK1) S114 Uniprot
O15530 (PDPK1) S241 Uniprot
O15530-4 (PDPK1) T386 Uniprot
O15530 (PDPK1) S393 Uniprot
O15530 (PDPK1) S396 Uniprot
O15530 (PDPK1) S410 Uniprot
O15530 (PDPK1) T513 Uniprot
O15530 (PDPK1) S529 Uniprot
P05106 (ITGB3) T779 Uniprot
P05771 (PRKCB) T500 Uniprot
P08559 (PDHA1) S232 Uniprot
P08559 (PDHA1) S293 Uniprot
P08559 (PDHA1) S300 Uniprot
P17612 (PRKACA) T198 Uniprot
P23443-2 (RPS6KB1) T229 Uniprot
P23443 (RPS6KB1) T252 Uniprot
P23443 (RPS6KB1) T412 Uniprot
P31749 (AKT1) T34 Uniprot
P31749 (AKT1) T308 Uniprot
P31749 (AKT1) S473 Uniprot
P31751 (AKT2) T309 Uniprot
P31751 (AKT2) S474 Uniprot
P36507 (MAP2K2) S222 Uniprot
P36507 (MAP2K2) S226 Uniprot
P51812 (RPS6KA3) S227 Uniprot
P51812 (RPS6KA3) S386 Uniprot
Q02156 (PRKCE) T566 Uniprot
Q02750 (MAP2K1) S218 Uniprot
Q02750 (MAP2K1) S222 Uniprot
Q05513-2 (PRKCZ) T227 Uniprot
Q05513 (PRKCZ) T410 Uniprot
Q05655 (PRKCD) T507 Uniprot
Q13153 (PAK1) T423 Uniprot
Q15349 (RPS6KA2) S218 Uniprot
Q15418 (RPS6KA1) S221 Uniprot
Q15418-2 (RPS6KA1) S230 Uniprot
Q16512 (PKN1) S773 Uniprot
Q16512 (PKN1) T774 Uniprot
Q16512-2 (PKN1) T780 Uniprot
Q16513 (PKN2) T816 Uniprot
Q16513-1 (PKN2) T951 Uniprot
Q96BR1 (SGK3) T320 Uniprot
Q96BR1-2 (SGK3) S454 Uniprot
Q96BR1-1 (SGK3) S486 Uniprot
Q99683 (MAP3K5) S966 Uniprot
Q99814 (EPAS1) T324 Uniprot
Q9HBY8-2 (SGK2) T193 Uniprot
Q9HBY8-1 (SGK2) T253 Uniprot
Q9HBY8-2 (SGK2) S356 Uniprot
Q9HBY8-1 (SGK2) S416 Uniprot
Q9UBS0 (RPS6KB2) T228 Uniprot
Q9UBS0-1 (RPS6KB2) S370 Uniprot
Q9UBS0-1 (RPS6KB2) T388 Uniprot
Q9Y243-2 (AKT3) T305 Uniprot

Research Backgrounds

Function:

Serine/threonine kinase which acts as a master kinase, phosphorylating and activating a subgroup of the AGC family of protein kinases. Its targets include: protein kinase B (PKB/AKT1, PKB/AKT2, PKB/AKT3), p70 ribosomal protein S6 kinase (RPS6KB1), p90 ribosomal protein S6 kinase (RPS6KA1, RPS6KA2 and RPS6KA3), cyclic AMP-dependent protein kinase (PRKACA), protein kinase C (PRKCD and PRKCZ), serum and glucocorticoid-inducible kinase (SGK1, SGK2 and SGK3), p21-activated kinase-1 (PAK1), protein kinase PKN (PKN1 and PKN2). Plays a central role in the transduction of signals from insulin by providing the activating phosphorylation to PKB/AKT1, thus propagating the signal to downstream targets controlling cell proliferation and survival, as well as glucose and amino acid uptake and storage. Negatively regulates the TGF-beta-induced signaling by: modulating the association of SMAD3 and SMAD7 with TGF-beta receptor, phosphorylating SMAD2, SMAD3, SMAD4 and SMAD7, preventing the nuclear translocation of SMAD3 and SMAD4 and the translocation of SMAD7 from the nucleus to the cytoplasm in response to TGF-beta. Activates PPARG transcriptional activity and promotes adipocyte differentiation. Activates the NF-kappa-B pathway via phosphorylation of IKKB. The tyrosine phosphorylated form is crucial for the regulation of focal adhesions by angiotensin II. Controls proliferation, survival, and growth of developing pancreatic cells. Participates in the regulation of Ca(2+) entry and Ca(2+)-activated K(+) channels of mast cells. Essential for the motility of vascular endothelial cells (ECs) and is involved in the regulation of their chemotaxis. Plays a critical role in cardiac homeostasis by serving as a dual effector for cell survival and beta-adrenergic response. Plays an important role during thymocyte development by regulating the expression of key nutrient receptors on the surface of pre-T cells and mediating Notch-induced cell growth and proliferative responses. Provides negative feedback inhibition to toll-like receptor-mediated NF-kappa-B activation in macrophages. Isoform 3 is catalytically inactive.

PTMs:

Phosphorylation on Ser-241 in the activation loop is required for full activity. PDPK1 itself can autophosphorylate Ser-241, leading to its own activation. Autophosphorylation is inhibited by the apoptotic C-terminus cleavage product of PKN2 (By similarity). Tyr-9 phosphorylation is critical for stabilization of both PDPK1 and the PDPK1/SRC complex via HSP90-mediated protection of PDPK1 degradation. Angiotensin II stimulates the tyrosine phosphorylation of PDPK1 in vascular smooth muscle in a calcium- and SRC-dependent manner. Phosphorylated on Tyr-9, Tyr-373 and Tyr-376 by INSR in response to insulin. Palmitate negatively regulates autophosphorylation at Ser-241 and palmitate-induced phosphorylation at Ser-529 and Ser-501 by PKC/PRKCQ negatively regulates its ability to phosphorylate PKB/AKT1. Phosphorylation at Thr-354 by MELK partially inhibits kinase activity, the inhibition is cooperatively enhanced by phosphorylation at Ser-394 and Ser-398 by MAP3K5.

Autophosphorylated; autophosphorylation is inhibited by the apoptotic C-terminus cleavage product of PKN2.

Monoubiquitinated in the kinase domain, deubiquitinated by USP4.

Subcellular Location:

Cytoplasm. Nucleus. Cell membrane>Peripheral membrane protein. Cell junction>Focal adhesion.
Note: Tyrosine phosphorylation seems to occur only at the cell membrane. Translocates to the cell membrane following insulin stimulation by a mechanism that involves binding to GRB14 and INSR. SRC and HSP90 promote its localization to the cell membrane. Its nuclear localization is dependent on its association with PTPN6 and its phosphorylation at Ser-396. Restricted to the nucleus in neuronal cells while in non-neuronal cells it is found in the cytoplasm. The Ser-241 phosphorylated form is distributed along the perinuclear region in neuronal cells while in non-neuronal cells it is found in both the nucleus and the cytoplasm. IGF1 transiently increases phosphorylation at Ser-241 of neuronal PDPK1, resulting in its translocation to other cellular compartments. The tyrosine-phosphorylated form colocalizes with PTK2B in focal adhesions after angiotensin II stimulation.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Appears to be expressed ubiquitously. The Tyr-9 phosphorylated form is markedly increased in diseased tissue compared with normal tissue from lung, liver, colon and breast.

Subunit Structure:

Homodimer in its autoinhibited state. Active as monomer. Interacts with NPRL2, PPARG, PAK1, PTK2B, GRB14, PKN1 (via C-terminus), STRAP and IKKB. The Tyr-9 phosphorylated form interacts with SRC, RASA1 and CRK (via their SH2 domains). Interacts with SGK3 in a phosphorylation-dependent manner. The tyrosine-phosphorylated form interacts with PTPN6. The Ser-241 phosphorylated form interacts with YWHAH and YWHAQ. Binds INSR in response to insulin. Interacts (via PH domain) with SMAD3, SMAD4 and SMAD7. Interacts with PKN2; the interaction stimulates PDPK1 autophosphorylation, its PI(3,4,5)P3-dependent kinase activity toward 'Ser-473' of AKT1 but also activates its kinase activity toward PRKCD and PRKCZ.

Family&Domains:

The PH domain plays a pivotal role in the localization and nuclear import of PDPK1 and is also essential for its homodimerization.

The PIF-pocket is a small lobe in the catalytic domain required by the enzyme for the binding to the hydrophobic motif of its substrates. It is an allosteric regulatory site that can accommodate small compounds acting as allosteric inhibitors.

Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PDPK1 subfamily.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Endocrine system > PPAR signaling pathway.

· Organismal Systems > Immune system > T cell receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Excretory system > Aldosterone-regulated sodium reabsorption.

References

1). Wu et al. Long non‑coding RNA maternally expressed gene 3 affects cell proliferation, apoptosis and migration by targeting the microRNA‑9‑5p/midkine axis and activating the phosphoinositide‑dependent kinase/AKT pathway in hepatocellular carcinoma. Oncology Letters 2021 May;21(5):345. (PubMed: 33747202) [IF=2.9]

Application: WB    Species: Human    Sample: HCC cells

Figure 5. Effect of lncRNA MEG3 on the expression of miR-9-5p, MDK, apoptosis and migration-related proteins, and PDK/AKT pathway activation. (A) Effect of lncRNA MEG3 on miR-9-5p expression. (B) Western blot images. (C) Relative protein expression of MDK, caspase-3, caspase-9 and MMP1 determined by western blotting. (D) Effect of lncRNA MEG3 on PDK/AKT signaling. Cells in the control group were untransfected cells (n=3). *P<0.01 compared with the lncRNA MEG3 NC group. lncRNA, long non-coding RNA; MEG3, maternally expressed gene 3 (MEG3); HCC, hepatocellular carcinoma; NC, negative control; miR, microRNA; MDK, Midkine; PDK1, phosphoinositide-dependent kinase 1, p, phosphorylated; MMP-1, matrix metalloproteinase 1.

2). LncRNA MEG3 affects cell proliferation, apoptosis and migration by targeting miR-9-5p/MDK axis and activating PDK/AKT pathway in hepatocellular carcinoma.

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